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Biomedical subjects

H Hashizume

Publications and source records attributed to H Hashizume.

At least 73 records · Page 4Linked to original sources

Effects of steroid and lipopolysaccharide on spontaneous resorption of herniated intervertebral discs. An experimental study in the rabbit.

STUDY DESIGN: Histologic examination was performed on autografted intervertebral disc materials of rabbit models, which were partially incised through a retroperitoneal approach at L1-L2 and grafted within the posterior epidural space at L4. OBJECTIVE: To evaluate whether the resorption process of the herniated intervertebral disc is influenced and controlled by treatments with medications. SUMMARY OF BACKGROUND DATA: Regarding resorption of herniated intervertebral discs, recent studies of magnetic resonance images and histologic investigations of surgically resected specimens in lumber disc herniation patients have been reported. It has been shown that inflammatory factors may play an important role in the mechanism of resorption of the herniated intervertebral disc. However, little is known about the origin of newly formed vessels and inflammatory cells detected in herniated disc specimens from patients. In this study, The resorption process of disc material grafted into the epidural space was observed in a rabbit model. METHODS: Thirty-six adult rabbits were used. The L1-L2 intervertebral disc was partially incised through a retroperitoneal approach. The harvested disc material, which contains the nucleus pulposus and the anulus fibrosus were placed into the posterior epidural space at L4 of the same rabbit. The animals were divided into control, and steroid groups. The control group received no treatment after surgery. In the lipopolysaccharide group, rabbits were injected 1 mg/kg into the peritoneum immediately and at 7 days after surgery. In the steroid group, rabbits were injected with 1 mg/kg betamethasone into the epidural space daily from 1 to 7 days after surgery. Rabbits of each group were killed for histologic examination at 1, 2, 4, and 8 weeks after surgery. RESULTS: At 1 and 2 weeks after surgery, inflammatory cells and newly formed vessels were more frequently observed in the lipopolysaccharide group than in the control and steroid groups. At 4 weeks after surgery, derangement and loosening of collagen fibers were also observed in the lipopolysaccharide group. At 8 weeks after surgery, fragmentation and partial disappearance of matrix were observed in the control and lipopolysaccharide groups. Most of the intervertebral discs were replaced by fibrous tissues in the lipopolysaccharide group. However, the matrix of the intervertebral disc almost remained. CONCLUSIONS: Autologous intervertebral disc material grafted into the epidural space was penetrated by newly formed vessels produced from the epidural fat tissue and resolved as the result of inflammatory reaction. Lipopolysaccharide accelerated the replacement of grafted intervertebral disc by fibrous tissue, which suggests the resorption of the disc in the epidural space of the rabbit, whereas high-dose steroid suppressed the replacement.

Animals↗

Three-dimensional cytoarchitecture of rat pulmonary venous walls: a light and scanning electron microscopic study.

The three-dimensional architecture of the rat pulmonary veins was studied by light microscopy (LM) and scanning electron microscopy (SEM). For LM, the left lungs were fixed with formalin, sectioned and immunostained with an anti-alpha-smooth muscle actin (alpha-SMA) antibody in addition to conventional staining. For SEM, the specimens were fixed with glutaraldehyde and immersed in 30% KOH solution for 8 min followed by treatment of collagenase solution for more than 5 h. By LM, the smooth muscle cells stained with anti-alpha-SMA showed discontinuous, periodical thickenings of circular bundles in the wall of the venules, but they became thin and continuous in the larger vessels (or veins) that had a cardiac muscle layer on the outside. Under SEM, the smooth muscle cells formed circular-oriented bundles at constant intervals along the venules less than 100 microm in diameter. These bundles had circumferential constrictions in the lumen. The cardiac muscle cells, which appeared in large pulmonary veins of more than 100 microm, ran in a circular or oblique direction and completely surrounded the vessel wall outside of the thin continuous layer of smooth muscle cells. The muscle arrangements were considered to play a significant role in the return blood flow in rat pulmonary veins.

Actins↗

Changes in the extracellular matrix on the surface of sintered bovine bone implanted in the femur of a rabbit: an immunohistochemical study.

The interface of implanted True Bone Ceramics (TBC; sintered bovine bone; Koken, Tokyo, Japan) was examined. In the primary experiment, TBC was implanted into the bone marrow of a rabbit's femur. The extracellular matrices (types I, II, and III collagens and fibronectin) of decalcified specimens collected 1-48 weeks postoperatively were immunohistochemically examined. Undecalcified sections collected 6 weeks postoperatively were used for line analyses of calcium and phosphorus, by a scanning electron microscope-electron probe microanalysis (SEM-EPMA) method. In a secondary experiment, TBC was implanted into an osteochondral defect of a femoral condyle, harvested 1-12 weeks postoperatively, and decalcified to examine the extracellular matrices at the interface. In the bone marrow in the early phase, TBC had absorbed quantities of fibronectin. Immature bone (containing both types I and III collagens) in direct apposition to the ceramic surface had matured (containing type I collagen alone) in the TBC pores. SEM-EPMA revealed the continuity of high levels of calcium and phosphorus at the TBC-bone interface. In the secondary experiment, enchondral ossification or fibrous tissue formation was observed near the articular surface. However, in the subchondral layer, direct bone formation was observed in the TBC pores. It was concluded that TBC has excellent bioactivity for inducing maturation of new bone matrix on porous surfaces.

Animals↗

A new approach to the development of anti-ischemic drugs: protective drugs against cell injury induced by lysophosphatidylcholine.

Recent studies have revealed that lysophosphatidylcholine (LPC) produces mechanical and metabolic derangements in perfused working rat hearts and Ca2+-overload in isolated cardiac myocytes. Thus, LPC possesses an ischemia-like effect on the heart. Therefore, a drug that possesses an anti-LPC action would protect or improve ischemia/reperfusion damage. We examined the effects of various anti-ischemic drugs on the Ca2+ overload induced by LPC. Our data suggest that a drug with high lipophilicity possesses a protective effect on cell injury induced by LPC, probably because of preservation of membrane integrity.

Adrenergic beta-Antagonists↗

Protective effect of prazosin on the hydrogen peroxide-induced derangements in the isolated perfused rat heart.

The effect of prazosin, an alpha1-selective adrenoceptor antagonist, on the hydrogen peroxide (H2O2)-induced mechanical and metabolic derangements was studied in the isolated rat heart, which was perfused aerobically by the Langendorffs technique at a constant flow rate and driven electrically. H2O2 (600 microM) produced both mechanical dysfunction (e.g., increase in the left ventricular end-diastolic pressure) and metabolic damage (e.g., decrease in the level of adenosine triphosphate) associated with lipid peroxidation (e.g., increase in the level of malondialdehyde). The H2O2-induced mechanical and metabolic derangements were attenuated by 2.5, 5, or 10 microM prazosin, and the increase in the level of malondialdehyde was attenuated by 5 or 10 microM prazosin. Nevertheless, prazosin had practically no effects on the mechanical function and energy metabolism of the H2O2-untreated normal heart at 2.5 or 5 microM, although it reduced the mechanical function at 10 microM. Prazosin was shown to have a hydroxyl radical scavenging effect. These results suggest that prazosin attenuates the H2O2-induced mechanical and metabolic derangements probably because of attenuation of the H2O2-induced lipid peroxidation in the heart.

Animals↗

Effect of nonsteroidal anti-inflammatory drugs on lipid peroxidation by hydroxyl radical.

1. Effects of acetaminophen and sodium salicylate on hydoxyl radical elimination were studied using an electron-spin-resonance spin-trapping method. The effects of these agents on lipid peroxidation of the human erythrocyte membrane were also investigated by the thiobarbituric acid method. 2. Acetaminophen and sodium salicylate depressed hydroxyl radical generated by Cu2+/H2O2. 3. Acetaminophen inhibited Cu2+/H2O2-dependent lipid peroxidation; however, sodium salicylate enhanced Cu2+/H2O2-dependent lipid peroxidation.

Acetaminophen↗

Low-voltage backscattered electron imaging of non-coated biological samples in a low-vacuum environment using a variable-pressure scanning electron microscope with a YAG-detector.

A variable-pressure scanning electron microscope (VP-SEM) equipped with a high-sensitive backscattered electron (BSE) detector of the YAG type was applied to studies of biological tissue samples. The rat kidney and trachea were fixed, dehydrated in ethanol, critical point-dried and examined in the VP-SEM under a specimen chamber pressure of 1 to 150 Pa. The high-resolution surface images of the non-coated specimens were obtained in a low-vacuum (1-20 Pa) environment at an accelerating voltage of 5 kV, while the images at 10-20 kV contained information beneath the surface of the specimens. The observation in the VP-SEM equipped with YAG detector in a low-vacuum (1-20 Pa) environment at low (3-5 kV) accelerating voltages is useful for the three-dimensional analysis of the surface morphology of biological non-coated samples.

Animals↗

Propofol improves functional and metabolic recovery in ischemic reperfused isolated rat hearts.

UNLABELLED: Propofol attenuates mechanical dysfunction, metabolic derangement, and lipid peroxidation by exogenous administration of H2O2 in the Langendorff rat heart. In this study, we examined the effects of propofol on mechanical and metabolic changes, as well as on lipid peroxidation induced by ischemia-reperfusion, in isolated, working rat hearts. Rat hearts (in control-modified Krebs-Henseleit bicarbonate buffer) were treated with two doses (25 microM and 50 microM) of propofol in an intralipid vehicle. In the first protocol, propofol was administered during the preischemic and reperfusion period, whereas in the second, it was only administered during the reperfusion period. Ischemia (15 min) decreased peak aortic pressure (PAOP), heart rate (HR), rate-pressure product (RPP), coronary flow (CF), and tissue concentrations of adenosine triphosphate (ATP) and creatine phosphate. After postischemic reperfusion (20 min), the CF and tissue concentration of ATP recovered incompletely; however, PAOP, HR, and RPP did not. Ischemia-reperfusion also increased the tissue concentration of malondialdehyde (MDA). In both protocols, both doses of propofol enhanced recovery of PAOP, HR, RPP, CF, and tissue concentration of ATP during reperfusion, and inhibited the tissue accumulation of MDA. These results indicate that propofol improves recovery of mechanical function and the energy state in ischemic reperfused isolated rat hearts, and the mechanism may involve the reduction of lipid peroxidation during postischemic reperfusion. IMPLICATIONS: We evaluated the possible cardioprotective effects of propofol in isolated, working rat hearts subjected to 15-min ischemia, followed by 20-min reperfusion. We observed that propofol attenuated mechanical dysfunction, metabolic derangement, and lipid peroxidation during reperfusion. This latter finding seems to be one mechanism for cardioprotective effects of propofol.

Adenine Nucleotides↗

Phospholipase A2 is not responsible for lysophosphatidylcholine-induced damage in cardiomyocytes.

Lysophosphatidylcholine (LPC) is known to increase the intracellular concentration of Ca2+ ([Ca2+]i), leading to cell damage. In the present study we examined whether LPC activates phospholipase A2 (PLA2) and whether the activation of PLA2 is responsible for the LPC-induced cell damage in isolated rat cardiomyocytes. LPC (15 microM) produced an increase in [Ca2+]i, a change in cell shape from rod to round, and the release of creatine kinase (CK) accompanied by a significant elevation of the cellular level of nonesterified fatty acids (NEFA), especially arachidonic acid. Three PLA2 inhibitors, 7, 7-dimethyl-(5Z,8Z)-eicosadienoic acid (DEDA), 3-(4-octadecylbenzoyl)acrylic acid (OBAA), and manoalide, attenuated the LPC-induced accumulation of unsaturated NEFA to a similar degree. Nevertheless, whereas both DEDA and OBAA attenuated the LPC-induced increase in [Ca2+]i, change in cell shape, and release of CK, manoalide attenuated none of them. In the Ca2+-free solution, LPC did not increase [Ca2+]i with significantly less accumulation of NEFA, but it changed the cell shape from rod to round and increased the release of CK. These results suggest that exogenous LPC increases the PLA2 activity, which, however, may not be responsible for the LPC-induced damage in cardiomyocytes.

Animals↗

Protective effects of ranolazine, a novel anti-ischemic drug, on the hydrogen peroxide-induced derangements in isolated, perfused rat heart: comparison with dichloroacetate.

The effect of ranolazine, a novel anti-ischemic drug that stimulates the activity of pyruvate dehydrogenase, on hydrogen peroxide (H2O2)-induced mechanical and metabolic derangements was studied in isolated rat heart and compared with that of dichloroacetate (DCA), an activator of pyruvate dehydrogenase. The heart was perfused aerobically by the Langendorff's technique at a constant flow and driven electrically. H2O2 (600 microM) decreased the left ventricular developed pressure and increased the left ventricular end-diastolic pressure (i.e., mechanical dysfunction), decreased the tissue level of adenosine triphosphate (i.e., metabolic derangement), and increased the tissue level of malondialdehyde (MDA) (i.e., lipid peroxidation). These mechanical and metabolic derangements induced by H2O2 were significantly attenuated by ranolazine (10 or 20 microM). On the other hand, DCA (1 mM) was ineffective in attenuating the H2O2-induced mechanical and metabolic derangements. Ranolazine, however, did not modify the tissue MDA level, which was increased by H2O2. In the normal (H2O2-untreated) heart, ranolazine did not alter the mechanical function and energy metabolism. These results demonstrate that ranolazine attenuates mechanical and metabolic derangements induced by H2O2. It is suggested that the protective action of ranolazine against the H2O2-induced derangements is due to neither the energy-sparing, DCA-like, nor anti-oxidant effects.

Acetanilides↗

Direct observation of t-butyl alcohol frozen and sublimated samples using low-vacuum scanning electron microscopy.

Frozen biological specimens in t-butyl alcohol were examined under a low-vacuum environment in a "wet SEM" or "variable pressure SEM (scanning electron microscope)" equipped with a cooling stage and highly sensitive backscattered detector of the YAG type. After fixation with glutaraldehyde and osmium tetroxide, rat tissue blocks (tracheae and kidneys), and cultured human carcinoma cells were dehydrated with a graded series of t-butyl alcohol. The specimens were directly frozen on the cooling stage at -10 degrees C, evacuated to 20 Pa in the specimen chamber, and observed by detecting backscattered electrons at accelerating voltages of 5-6 kV. The images became clearer 20 min after the vacuum reached 20 Pa and revealed had good quality by 30 min, probably because t-butyl alcohol was sublimated during the time. The cilia of tracheal ciliated cells, end-feet of the podocytes of the renal glomerulus, and processes of cultured cells were clearly observed without any serious preparation artifacts. Since the low-vacuum SEM of t-butyl alcohol frozen samples is both simple and provides high imaging quality, it is expected to be useful in a variety of biological fields such as the rapid pathological diagnosis.

Animals↗

Fibrous structure and connection surrounding the metacarpophalangeal joint.

The fibrous components of the metacarpophalangeal (MP) joint including the palmar plate, the collateral ligament and the dorsal plate were studied with particular attention paid to the fibrous structure of the fibrous tendon sheath and the deep transverse metacarpal ligament. The tough fibrillar structure around the MP joint, especially the force nucleus, consisted of three types of mixed fibers: the fibrous tendon sheath of the A1 pulley, the deep transverse metacarpal ligament, and the palmar plate. The tendon sheath was located on the ulnar side in the index and middle fingers, on the central position in the ring finger, and on the radial side in the little finger. These fibrous connections among the fingers formed a transverse arch in the hand. The palmar plate of the MP joint was relatively rigid and appears to function as a cushion when flexed. A fold-like protrusion of the synovial layer of the palmar plate of the MP joint had a meniscoid function, which was larger than that of the proximal interphalangeal joint. The capsule of the MP joint was thicker at the dorsal area, forming a dorsal plate, which is a sliding floor of the extensor mechanism and has a meniscoid function for joint congruity. The main lateral stabilizer consisted of collateral ligaments and accessory collateral ligaments anchored to the palmar plate. These structures act together as a "phalangeal cuff", connecting the proximal phalanx to the metacarpal head and stabilizing the MP joint.

Aged↗

Possible mechanism of painful radiculopathy in lumbar disc herniation.

The pathophysiologic mechanisms of painful radiculopathy caused by a herniated intervertebral disc remain unknown. This study sought to determine whether the autologous intervertebral disc produces pain related behavior and whether phospholipase A2 and nitric oxide are involved in the pathophysiologic mechanism producing the behavior. A rat model, in which autologous intervertebral discs were implanted on the nerve root in the lumbar spine, was used to measure hyperalgesia, which is a pain related behavior in the rat. In this experimental model, autologous nucleus pulposus and anulus fibrosus transplanted to lumbar nerve roots produced mechanical and thermal hyperalgesia, respectively. Epidural injection of a selective inhibitor for phospholipase A2 resulted in the disappearance of hypersensitivity to noxious mechanical stimuli. Thermal hyperalgesia produced by application of the anulus fibrosus was abated and abolished by epidural injections of saline and one of the inhibitors for nitric oxide synthase, respectively. The authors suggest that chemical mediators such as phospholipase A2 and nitric oxide, induced by extruded or sequestrated intervertebral discs, are involved in the pathophysiologic mechanisms of painful radiculopathy in lumbar disc herniations. This study may be useful in attempting to develop new medical approaches for treatments of lumbar disc herniation.

Analysis of Variance↗

[A case report of dumbbell neurogenic tumor of the superior mediastinum].

A 15-year-old female was admitted because of the superior mediastinum mass on chest X-rays and sensory loss of ulnar side of the left arm. Computed tomographic scanning and magnetic resonance imaging revealed that the tumor was dumbbell-shaped and invaded the vertebral canal through the intervertebral foramen between C 7 and Th 1. At first laminectomy of vertebrae (C 6-Th 1) was made in a prone position and intra-spinal portion of the tumor was resected. Then the patient was placed in a supine position and the chest was opened by left hemicollar incision and sternotomy to the 2nd intercostal space. The tumor was divided into two parts at the level of 1st rib and completely removed. The pathological diagnosis was schwannoma. This procedure is safe and useful for dumbbell type tumor located in superior mediastinum, especially in case of large tumor from neck to the thoracic cavity.

Adolescent↗

Differential effects of Ca2+ channel blockers on Ca2+ overload induced by lysophosphatidylcholine in cardiomyocytes.

The effects of Ca2+ channel blockers (verapamil, diltiazem, nicardipine, bepridil and flunarizine) on Ca2+ overload induced by lysophosphatidylcholine were examined in rat isolated cardiomyocytes. Addition of lysophosphatidylcholine (15 microM) produced Ca2+ overload as evidenced by a marked increase in the concentration of intracellular Ca2+ and hypercontracture of cells. Verapamil, flunarizine and bepridil concentration dependently inhibited the lysophosphatidylcholine-induced Ca2+ overload, whereas diltiazem and nicardipine did not. Lysophosphatidylcholine increased the release of creatine kinase, which was significantly attenuated by verapamil, flunarizine or bepridil (5 microM for each), but not by diltiazem or nicardipine (20 microM for each). Verapamil, flunarizine, bepridil (which attenuated the lysophosphatidylcholine-induced Ca2+ overload) and nicardipine (which did not) inhibited the veratridine-induced increase in the concentration of intracellular Na+ (indicated by the increase in fluorescence ratio of Na(+)-binding benzofuran isophthalate) and cell contracture, whereas diltiazem did not. These results suggest that verapamil, bepridil and flunarizine attenuate the Ca2+ overload induced by lysophosphatidylcholine, and that the Ca2+ channel blocking action of these drugs does not contribute substantially to this effect. The Na+ channel inhibition together with high lipophilicity of these drugs may be important for the attenuation of the lysophosphatidylcholine-induced Ca2+ overload.

Animals↗

The role of phospholipase A2 and nitric oxide in pain-related behavior produced by an allograft of intervertebral disc material to the sciatic nerve of the rat.

STUDY DESIGN: To elucidate the pathomechanisms of radicular pain secondary to lumbar disc herniation. OBJECTIVES: To evaluate whether intervertebral disc material applied to the sciatic nerve produces hyperalgesia, and if the hyperalgesia in influenced by inhibitors of phospholipase A2 and nitric oxide synthase. SUMMARY OF BACKGROUND DATA: Previously, the authors reported that application of nucleus pulposus and anulus fibrosus material to the lumbar epidural space produces different forms of hyperalgesia (mechanical versus thermal), with different and distinct histologic changes. Additional pharmacologic studies showed that phospholipase A2 and nitric oxide are involved in the mechanisms that produce the mechanical and thermal hyperalgesia, respectively, N omega-nitro-L-arginine methyl ester and mepacrine are relatively selective inhibitors of nitric oxide synthase and phospholipase A2, respectively. However, it is not known what the relation is between the hyperalgesia produced and the activation and involvement of phospholipase A2 and production of nitric oxide, or why the application of nucleus pulposus and nucleus pulposus with anulus fibrosus produces different types of hyperalgesia. METHODS: Experiments were performed in five groups of rats: The control group (no treatment), the sham group (exposure of the sciatic nerve only), the fat group (allografted fat on the sciatic nerve), the nucleus pulposus group (allografted nucleus pulposus) and the nucleus pulposus + anulus fibrosus group (allografted nucleus pulposus and anulus fibrosus). Withdrawal threshold and latency from mechanical pressure and a radiant heat to hind paws were measured preoperatively and postoperatively. After local sciatic nerve administration of N theta-nitro-L-arginine methyl ester or mepacrine into the operated site, sensitivities to noxious stimuli were reevaluated after treatment. RESULTS: Only rats in the nucleus pulposus group showed evidence of mechanical hyperalgesia. However, injection of N theta-nitro-L-arginine methyl ester resulted in evidence of mechanical hyperalgesia in the nucleus pulposus + anulus fibrosus group. Mechanical hyperalgesia was produced in the nucleus pulposus group and after injection of N theta-nitro-L-arginine methyl ester in the nucleus pulposus+anulus fibrosus group, both of which returned to normal after mepacrine injection. There were no significant changes in sensitivity to thermal stimuli in any of the experimental groups. CONCLUSION: It appears that phospholipase A2 and nitric oxide play important but different roles in pathomechanisms of radicular pain in lumbar disc herniation.

Animals↗

Histochemical demonstration of nitric oxide in herniated lumbar discs. A clinical and animal model study.

STUDY DESIGN: This study was designed to localize the cells that produce nitric oxide in a lumbar disc herniation by histochemical method, including in situ hybridization. OBJECTIVE: To clarify which cells in herniated lumbar discs produce nitric oxide. SUMMARY OF BACKGROUND DATA: It was reported that herniated lumbar intervertebral disc specimens in culture are capable of producing nitric oxide. METHODS: Surgical specimens from lumbar disc herniation were examined to determine nitric oxide synthase histologically using nicotinamide adenine dinucleotide phosphate diaphorase histochemistry. Allografts of intervertebral disc materials were placed on the epidural space at L6 level in the rat. Nitric oxide synthase was examined in the applied tissues using nicotinamide adenine dinucleotide phosphate diaphorase histochemistry and in situ hybridization histochemistry. RESULTS: Nicotinamide adenine dinucleotide phosphate diaphorase (nitric oxide synthase) positive cells were observed in 2 (40%) of 5 herniated disc materials in patients. The positive cells were mainly in granulation tissue around intervertebral disc materials. In animal models, nitric oxide synthase-positive cells were observed in all specimens at 1 and 2 weeks postoperatively. Newly formed vessels and small round cells in granulation tissue around the grafted intervertebral disc showed positive reaction. In situ hybridization demonstrated the expression of inducible isoform of nitric oxide synthase messenger RNA (mRNA) identical to small round cells around the applied intervertebral disc. CONCLUSION: Nitric oxide in a lumbar disc herniation is mainly produced by cells in granulation tissue around the herniated intervertebral disc.

Adult↗